Objective To investigate the traditional Chinese medicine ( TCM) syndromes of rheumatoid arthritis ( RA) by studying the Chinese medical patterns,and analyze the correlations of different indicators and related syndromes. Methods The questionnaires were designed according to clinical epidemiological theories. The data were collected and analyzed by multivariate statistical methods. Results The TCM symptoms of the 401 cases of RA were concluded into 3 common factors,that is,cold,heat and deficient. Their syndromes were sub-classified as 6 groups: cold syndrome,heat syndrome,deficiency,cold heat complex,deficient cold and asthenic fever. Statistical analysis showed that there were significant differences in health assessment questionnaire ( HAQ) ,platelet ( PLT) ,rheumatoid factor ( RF) ,anti-keratin antibody ( AKA) among these 6 TCM syndromes ( P

Objective To observe the effects of S100A4 siRNA on the expression of serum TNF‐α,IL‐1βand VEGF in adjuvant arthritis rats .Methods Adjuvant arthritis rat models were established and were randomly divided into model group and interfere group .On Day 11 ,rats in interfere group were injected with S100A4 siRNA fragment in articular cavity .Arthritis index (AI) chan‐ges and pathological changes of ankle joint were observed .The levels of serum TNF‐α and IL‐1β ,VEGF were detected by ELISA . Results Compared with that of model group ,the levels of serum TNF‐ α ,IL‐1β and VEGF were reduced significantly in interfere group (P< 0 .05) ;variances of AI and pathological scores in interfere group were diminished significantly (P< 0 .05) .Conclusion Inhibition of the expression of S100A4 gene can significantly reduce the expression of inflammatory factor TNF‐α ,IL‐1 β and angio‐genesis factor VEGF ,and improve the pathological injury of synovial membrane .

BACKGROUND: Sinomenine is an alkaloid monomer extracted from a Chinese medicinal herb sinomenium acutum stem. It is used in the therapy of the rheumatoid arthritis and has clear and definite therapeutic effects, but the therapeutic mechanism is unclear.OBJECTIVE: To observe the effect of sinomenine at different doses in vitro on the activity of nuclear factor-κB(NF-κβ) and mRNA expressions of the tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) andinterleukin-10 (IL-10) in the synovial cells of the rats with adjuvant arthritis(AA) to explore the probable mechanism of sinomenine in the treatment of rheumatoid arthritis(RA).DESIGN: A controlled repeated measuring study based on the cells.SETTING: Department of traditional chinese medicine and the institute of burn research of a military medical university.MATERIALS: This study was finished at the Laboratory of the Institute of Burn Research of Chinese PLA. The experimental animals were 25 healthy male Wistar rats of clean grade. The AA model rats were made and the synovial cells were collected and grouped as follows: normal control group, AA group,AA + sinomenine 30 mg/L group, AA + sinomenine 60 mg/L group, AA + sinomenine 120 mg/L group. The activity of the NF-κB was measured by the electrophoresis mobility shift assay(EMSA) . The mRNA expressions of the TNF-α, IL-1β and IL-10 were measured by reverse transcription-PCR assay.MAIN OUTCOME MEASURES: The results of the changes of the activity of the NF-κB and the mRNA expressions of the TNF-α, IL-1β and IL-10 in the synovial cells of the rats with adjuvant arthritis after the treatment with sinomenine at different doses.RESULTS: Compared with the normal control group, the activity of the NF-κB and the mRNA expressions of the TNF-α, IL-1β and IL-10 in the synovial cells in the AA group all increased significantly and the outcomes were 17±6, 0.570±0.047, 0.730±0.093, 0. 683 ±0.081 (t= 2.71 -4.07, P < 0.05). After the administration of sinomenine, the activity of NF-κB showed a good correlation with mRNA expressions of the TNF-αandIL-13(r=0.810, P <0.001; r=0.562, P <0.05), but no statistical relevance with mRNA expression of IL-10 was established. Sinomenine showed a dose-dependent inhibition on the activity of the NF-κB and the mRNA expressions of the TNF-α and IL-1β in a certain range of concentrations(30-120 mg/L), but no dose-dependent inhibition on mRNA expression of the IL-10 was observed.CONCLUSION: Through the inhibition of the activity of the NF-κB,sinomenine decreased the mRNA expressions of the TNF-α and the IL-1β in the synovial membrane cells.

[ ABSTRACT] AIM:To explore the role of SHARPIN in regulation of Rip1 in castration-resistant prostate cancer LNCaP-AI cells.METHODS:The LNCaP-AI cells were treated with TNF-α+Z-VAD ( an inhibitor of pan-caspase) to activate necroptosis, which were compared to the cells treated with TNF-α+Z-VAD+Nec-1 ( an inhibitor of Rip1 ) .A blank group and a TNF-α-treated group were set up as controls.The cell viability in each group was measured by MTS as-say.In addition, SHARPIN was knocked down by siRNA, and the inhibitory efficiency was evaluated by RT-qPCR.The expression of Rip1 at mRNA and protein levels after knocking down SHARPIN was determined by RT-qPCR and Western blot to explore the underlying mechanism of regulatory network of necroptosis in prostate cancer.RESULTS: Compared with blank control group and TNF-α-treated group, the viability of LNCaP-AI cells treated with TNF-α+Z-VAD decreased by 28%(P<0.05).After treated with TNF-α+Z-VAD+Nec-1, the LNCaP-AI cells showed no significant difference in the viability compared with blank control and TNF-α-treated groups.Taken together, necroptosis may be an important way of cell death in LNCaP-AI cells.Besides, the expression of Rip1 at protein level was up-regulated following the inhibition of SHARPIN using siRNA, indicating that down-regulation of SHARPIN enhanced necroptosis via activating Rip1 in
LNCaP-AI cells.CONCLUSION:Necroptosis is an important way of cell death .Inhibition of oncogenic factor SHARPIN enhances necroptosis via activating Rip1 in LNCaP-AI cells.

Objective To construct a bait vector containing human Foxp3 gene in yeast two-hybrid system in order to screen the cDNA library of T lymphocyte. Methods RT-PCR was used to amplify the Foxp3 gene fragment from the peripheral blood mononuclear cells (PBMC) with the primers designed in accordance with the sequence in GenBank. The product was inserted into pMD18-T vector. After verified with restriction endonuclease digestion of EcoRⅠ and SalⅠ, the vector was inserted into the “bait plasmid” pGBKT7 (named as pGBKT7-Foxp3). After confirmation with restricted endonuclease digestion and sequence analysis, the plasmid was transformed into the yeast cell AH109, and its toxicity and transcriptional activation was tested by both the phenotype assay and the color assay. Results The amplified product of 1 203 bp was inserted into PMD18-T vector and proven correctly by double restriction enzyme digestion. Sequence analysis revealed that the fragment was correctly inserted into pGBKT7 with a right reading frame and its expression in yeast was verified. Conclusion The bait plasmid pGBKT7-Foxp3 constructed expresses correctly, and can not activate the transcription of reporter gene alone in yeast two-hybrid system

[ ABSTRACT] AIM: To investigate the SALL4 expression, proliferation and apoptosis in the LNCaP cells after transfection of SALL4 siRNA.METHODS: The expression of SALL4 at mRNA and protein levels was detected by real-time PCR and Western blotting.MTS assay, colony formation assay and flow cytometry were used to determine the prolifer-ation, colony formation ability and apoptosis of the LNCaP cells.The effect of SALL4 on the expression of Bax and Bcl-2 was analyzed by Western blotting.RESULTS:Compared with negative control group, the expression of SALL4 at mRNA and protein levels in LNCaP cells was down-regulated by transfection of SALL4 siRNA ( P<0.05 ) .The proliferation rate and colony formation ability were decreased, while apoptosis rate increased in si-SALL4 group (P<0.05).Higher expres-sion of Bax and lower expression of Bcl-2 in si-SALL4 group were observed ( P<0.05 ) .CONCLUSION:Down-regula-tion of SALL4 by siRNA not only suppresses LNCaP cell proliferation and colony formation, but also inhibits Bcl-2 expres-sion and activates Bax expression to induce apoptosis.

Objective To establish the methodology of combining BOLD and 1H-MRS for investigating correlation between the deactivation in medial prefrontal cortex (MPFC) and gamma-aminobutyric acid (GABA) concentration by acupuncture at LI4 (Point Hegu),and to optimize the experimental technique and procedure.Methods Twenty healthy adult volunteers were enrolled.During fMRI-BOLD scanning,each subject received acupuncture at right LI4 (Point Hegu).MRS scanning was based on MEGA-PRESS sequence,and ROIs were located at bilateral MPFC.The task BOLD fMRI was block design,including 3 stimulations (30 s) with 2 intervals (2 min).Then MRS scanning was performed before and after BOLD.The quantitative values of the BOLD positive and negative activations (Pm) and GABA concentrations were calculated.Results All 20 subjects completed BOLD fMRI scanning,and met the postprocessing requirements.MRS images of 9 subjects with good image quality were included in analysis.Among all 20 subjects,positive activation (Pm=1.17± 0.16) was observed in 9,while negative activation (Pm =-1.31 ± 0.17) was observed in 11 subjects.The GABA average values before and after the acupuncture were (19.93 ±1.04) nmol/L and (20.04±0.81)nmol/L,respectively,and the average amplitude between post-and pre-acupuncture was (0.11 ± 1.60)nmol/L.Conclusion The success rate of this method for quantitative study of brain function established multimodal-functional (BOLD-fMRI and MRS) was acceptable,and the multimodal brain function changes as well as the quantitative values were observed in the brain region during acupuncture.Combined BOLD and MRS quantitative method is feasible for testing acupuncture response in the brain.

Methyltransferase like 13 (METTL13), a kind of methyltransferase, is implicated in protein binding and synthesis. The upregulation of METTL13 has been reported in a variety of tumors. However, little was known about its potential function in head and neck squamous cell carcinoma (HNSCC) so far. In this study, we found that METTL13 was significantly upregulated in HNSCC at both mRNA and protein level. Increased METTL13 was negatively associated with clinical prognosis. And METTL13 markedly affected HNSCC cellular phenotypes in vivo and vitro. Further mechanism study revealed that METTL13 could regulate EMT signaling pathway by mediating enhancing translation efficiency of Snail, the key transcription factor in EMT, hence regulating the progression of EMT. Furthermore, Snail was verified to mediate METTL13-induced HNSCC cell malignant phenotypes. Altogether, our study had revealed the oncogenic role of METTL13 in HNSCC, and provided a potential therapeutic strategy.
Head and Neck Neoplasms ; Humans ; Squamous Cell Carcinoma of Head and Neck/genetics*